Cell and Developmental Biology Commons^™

The Development Of An In Vivo Microdialysis Collection Method Of Cytokines From Brain Tissue, Anthony W. Herbaugh

Graduate Theses and Dissertations

In this thesis, different methods to improve the microdialysis collection procedure for cytokines from brain tissue are presented. The first method was based on stopped flow and results indicating that no significant difference in relative recovery between stopped flow and continuous flow are shown. The second method is an antibody bead-based enhancement method. With the antibody bead-based method, a 3.5 fold increase in the collected concentrations of Chemokine (C-C motif) Ligand 2 (CCL2) were observed. However, there was no significant increase in the in vivo collection efficiency of Interleukin-6 (IL-6) using the antibody enhancement. Finally the development of an in-house …

Proteomic And Biochemical Studies Of Estrogen-Mediated Signaling And Novel Estrogen Receptor-Interacting Proteins In Breast Cancer Cells, Zhenqi Zhou

Graduate Theses and Dissertations

Estrogen plays essential roles in the growth, development, and homeostasis of a number of tissues, and can also be linked to the growth of breast cancer. The biological activities of estrogen are mediated by estrogen receptors (ERs) ERá and ERâ, and also orphan G-protein-coupled receptor 30 (GPR30). In order to identify novel proteins that are involved in ER-mediated actions of estrogen, we used mass spectrometry-based quantitative proteomic methods to systematically profile global protein expression in responses to E2 (17â-estradiol) stimulation in human breast cancer cell, and identify and characterize cellular novel proteins that are associated with ERs in breast cancer …

Expansion Of Human Induced Pluripotent Stem Cells On Synthetic Substrate In Defined Medium, Huantong Yao

Graduate Theses and Dissertations

Human induced pluripotent stem cells (hiPSCs) have the potential to generate patient-specific cells to treat many incurable diseases by cell replacement therapy. However, so far the culture of hiPSCs depends greatly on feeder cells or Matrigel which has safety issues. Thus, chemically defined substrates that could provide niches necessary for cell attachment and proliferation are preferred for clinical application of hiPSCs. Recently, Corning Life Sciences has developed synthetic peptide-functionalized cell culture surface, referred to as Corning® Synthemax that support self-renewal and differentiation of human embryonic stem cell (hESC). In this work, we have collaborated with Corning to investigate the attachment, …

Phenotypic Plasticity Of Rattlesnake Trophic Morphology, Matthew Thomas Smith

Graduate Theses and Dissertations

The trophic morphology of gape-limited predators constrains the shape and size of prey items they can ingest. Trophic morphology consists of any morphological feature that is involved in the handling and ingestion of food. Diet has a profound effect on the morphology of many gape-limited predators. Identifying how prey type and resource level affect the morphology of different populations is an essential step in understanding the mechanisms contributing to patterns of morphological diversity. Species interactions (Chapter 1) induce plasticity in morphology that can lead to increased fitness, morphological divergence, and eventually speciation.

In Chapter 2, a laboratory study tested the …

Cell Bioenergetics In Leghorn Male Hepatoma Cells And Immortalized Chicken Liver Cells In Response To 4-Hydroxynonenal Induced Oxidative Stress, Alissa Laura Piekarski

Graduate Theses and Dissertations

Bioenergetic mechanisms responsible for ATP production are essential in carrying out maintenance and cell-specific functions. In this study, hepatocytes (liver cells) were used to test both endogenous and exogenous stress on cellular respiration. The secondary lipid peroxide, 4-hydroxynonenal (HNE), was used because it alters bioenergetics by increasing mitochondrial proton leak that attenuates mitochondrial radical production and, therefore, endogenous oxidative stress. The major objective of this study was to demonstrate effects of HNE-induced oxidative stress on avian hepatocyte bioenergetics. Various chemical which help enable the determination of oxygen (O2) consumption linked to ATP synthesis (oligomycin), maximal O2 consumption (FCCP), and proton …

Peptoid Based Slide Coatings For Disease Detection Via Elisa Microarray Analysis, Melissa Lea Hebert

Graduate Theses and Dissertations

Poly-N-substituted glycines (peptoids) are a very versatile family of synthetic molecules that can be customized for any number of applications. In this study, we chose to use peptoids as a foundation for sandwich ELISA microarray analysis with a long term goal of creating an early detection device for complex diseases such as cancer. The peptoids were designed to self-assemble into microspheres to be used in coatings on the surface of the microarray substrates to increase the surface area available for antibody attachment. This increased antibody density would lead to an increase in the microarray analysis sensitivity and dynamic range. Studies …

An Analysis Of The Redox Properties And Stability Of Chlamydomonas Reinhardtii Cytochrome F, Cytochrome C6, And Mutants Thereof, Nicole Lynn Vanderbush

Graduate Theses and Dissertations

This body of work presents mutagenesis studies conducted on two c-type cytochromes from Chlamydomonas reinhardtii. Cytochrome f, a unique c-type cytochrome, is investigated in regards to its redox potential, the dependence of the redox potential, and the thermal stability of the protein. The mutations made were Y1F, Y9F, Y160F, Y160L, R156L, and R156K. The residues that were mutated surround the heme. It was found that, relative to the wild-type, only the Y160L and R156 mutants showed any difference in midpoint potential at pH 7. Wild-type and mutants both had a midpoint potential that was dependent upon pH indicating that none …

Interactions Of Nitric Oxide And Superoxide Pathways In Hyperglycemic Endothelial Cells, Steven Clay Rogers

Graduate Theses and Dissertations

Cardiovascular complications arising from diabetic hyperglycemia represents one of the leading causes of death and greatest public health challenges of modern societies. Despite state-of-the-art glucose control, diabetic patients remain at a markedly increased risk of cardiovascular disease. The loss of endothelial function (the development of diabetic endothelial dysfunction) has been implicated in the development of numerous diabetic cardiovascular diseases. The endothelial cell produces many vasoactive substances, hormones and cytoprotective biological factors. Endothelial cells are also involved in and affected by the initiation of inflammatory responses through the release and interaction of cytokines and other immune system molecules. Therefore, regulation of …

Electrospun Polycaprolactone Nanofiber Scaffolds For Tissue Engineering, Andreas Haukas

Graduate Theses and Dissertations

Stem cell and tissue engineering offer us with a unique opportunity to research and develop new therapies for treating various diseases that are otherwise incurable using traditional medicines. However, development of these new therapies replies upon the establishment of in vitro cell culture and differentiation systems that mimic in vivo microenvironments required for cell-cell and cell-ECM interaction. The development of these cell culture systems depends upon the identification of appropriate biomaterials and cell sources. Biomaterials should be carefully selected and fabricated into scaffolds for supporting cell growth and differentiation. In this study, we explored the fabrication of 3D electrospun nanofiber …

Genome-Wide Identification Of Conditionally Essential Genes In Salmonella Typhimurium Using Tn-Seq Method, Anita Khatiwara

Graduate Theses and Dissertations

ABSTRACT

As more whole genome sequences become available, there is an increasing need for high-throughput methods that link genes to phenotypes and facilitate discovery of new gene functions. The objective of this study was to develop a high-throughput method to study gene functions in bacteria and use this method to study gene functions of S. enterica serotype Typhimurium (S. Typhimurium) under various environmental conditions encountered during its life cycle. Chapter I of this dissertation reviews the history and evolution of functional genomics in bacteria with focus on Salmonella, along with the recent techniques available. Chapter II, deals with the development …

In Vitro Selection Of Aptamers Against Avian Influenza Virus H5n1, Jingjing Zhao

Graduate Theses and Dissertations

Over $10 billion losses in the poultry industry were caused by avian influenza (AI) so far. Rapid and specific detection of avian influenza virus is urgently needed with the concerns over the outbreaks of highly pathogenic H5N1 influenza virus and cases of animal and human infection. Aptamers are oligonucleic acid or peptide molecules that bind a specific target molecule with good affinity. They show better thermal stability than antibodies. The goal of this research was to select DNA-aptamers as the specific recognition element of AI H5N1virus to be used in detection assays specific for field application. In this study, Systematic …

Mechanism Of Transcriptional Suppression Of A Phytochrome A Epiallele In Arabidopsis Thaliana, Gulab D. Rangani

Graduate Theses and Dissertations

Cytosine methylation in DNA is an integral part of epigenetically controlled regulatory networks in eukaryotes. Both plants and vertebrates display DNA methylation in the gene coding region; however, its role in gene expression is not well understood. Gene promoter, on the other hand, remains largely unmethylated. Acquisition of methylation in promoter results in transcriptional suppression of the gene. The goal of this research is to study the effect of coding region methylation in gene expression using a unique gene model, phyA'. phyA' is a transcriptionally suppressed epiallele of the Arabidopsis thaliana Phytochrome A gene, which contains methylation in CG sites …

Effects Of Lactate Dehydrogenase Haplotypes And Body Condition On Beef Cow Production, Olfat Taleb Alaamri

Graduate Theses and Dissertations

Lactate dehydrogenase (LDH) catalyzes the conversion of the pyruvate to lactate (forward) or lactate to pyruvate (reverse) in the last step of glycolysis. Objectives were to document the effects of LDH haplotypes and its SNPs, found in the promoter and coding sequence site, and body condition on beef cow production. Four single nucleotide polymorphisms (SNP) of LDH-B and Five single nucleotide polymorphisms of LDH-A were detected. Eight haplotypes of LDH-B were assigned with the same order of SNPs: G-348A, A-261G, N-222D, and C541A and four haplotypes of LDH-A were assigned with the same order of SNPs: T-327G, D-263C, G390A, A406G, …

The Role Of Multiple Ccaat-Binding Factors In Candida Albicans Gene Expression, Lashall Lynn Bates

Graduate Theses and Dissertations

The CCAAT-binding factor is a heterooligomeric transcription factor that is evolutionarily conserved in eukaryotes. In yeast, the DNA-binding component that interacts with the CCAAT consensus sequence in promoters consists of the subunits termed Hap2p, Hap3p and Hap5p. In yeast and fungi, a fourth subunit, Hap4p, is required for regulating gene expression. The goal of this research is to understand the function of the Candida albicans CCAAT-binding factor and how it relates to virulence and pathogenicity. C. albicans is a human opportunistic pathogen responsible for a variety of mucosal and systemic infections that result in significant morbidity and mortality, particularly in …

Characterization And Functional Regulation Of Bioactive Peptides In Avian Macrophages And Heterophils, Lakshmi Kannan

Graduate Theses and Dissertations

Oligopeptides and low molecular weight polypeptides play central roles as effectors and signal transducers acting as hormones, neurotransmitters, growth factors, toxins, and antimicrobial factors that are important for the survival of the organism. Owing to the ubiquitous involvement of peptides in many key regulatory processes, we have been interested to identify native peptides in different cells and tissues and understand their functions. To conduct our studies, we used avian macrophages and heterophils as models of specialized cells which constitute central components of innate immunity. These studies involved (a) qualitative identification and characterization of the peptides associated with high intensity mass …

The Membrane Interface Of Chloroplast Signal Recognition Particle-Dependent Protein Targeting, Naomi Jane Marty

Graduate Theses and Dissertations

A novel signal recognition particle (SRP) found in the chloroplast (cpSRP) works in combination with the cpSRP receptor, cpFtsY, to facilitate the post-translational targeting of a family of nuclear-encoded thylakoid proteins to the Alb3 translocase in thylakoid membranes. Work here focused on understanding events at the membrane that take place to ensure targeting of the cpSRP-dependent substrate to Alb3. Specifically, we sought to understand the structural and functional role of membrane binding by cpFtsY, a protein that exhibits the ability to partition between the membrane (thylakoid) and soluble (stroma) phase during protein targeting. We also sought to understand whether a …

Preparation, Separation, Purification, Characterization And Human Cell Line Anti-Cancer Evaluation Of Rice Bran Peptides, Arvind Kannan

Graduate Theses and Dissertations

Bioactive compounds are revolutionizing the nutritional and medicinal world with their inherent disease-fighting properties. A wide range of functional groups fall under the category of imparting health benefits. Compounds from both animal and plant origins have been generated as bioactive agents that have opened up new vistas for alternative medicine and natural healing. For example, in a debilitating disease like cancer, these compounds can act to suppress or delay the underlying pathology over and above the conventional treatment strategies involving drugs or chemotherapy. In other words conventional and invasive therapy, although still considered appropriate at certain stages of cancer, their …